Funding is requested for a Special Order Research Product BD FACSAria II cell sorter for the UCLA Flow Cytometry Core Facility (FCCF) established in 1988. Last year, under Director Dr. Beth Jamieson and Manager Ingrid Schmid, the FCCF provided sorting for 80 UCLA laboratories. Direct grants from the Jonsson Cancer Center and the UCLA AIDS Institute, the David Geffen School of Medicine, grants from the UCLA Chancellor's office, and user fees support the FCCF. The BD Aria II will replace a FACSVantage SE that is over 8 years old and lacks the technical capabilities required by FCCF users. These shortcomings have created scheduling conflicts for access to the single BD FACSAria currently in the FCCF. Compared to the FACSVantage, the FACSAria II will be equipped with: enhanced high-speed sorting and electronics for increased sort recoveries, higher fluorescence sensitivity, a yellow-green laser for improved detection of PE and PE- tandem fluorochromes, 10 fluorescence detectors, improved safety features for biohazardous sorts, and the Automated Cell Deposition Unit for single cell cloning. These are features UCLA investigators have indicated as critical for the progress of their research. To meet the very high demand for timely cell sorting, the FCCF has maintained three cell sorters since 2003. However, two are FACSVantage sorters which are technologically outdated and will be losing manufacturer support for service and parts. The one FACSVantage we are asking to replace through this grant was used by 41 NIH-funded investigators in the last year alone. Should the instrument break down, it would jeopardize the many research projects that depend on it for cell separation. The new BD FACSAria II will relieve the deficiencies of the current instruments in the FCCF, offer state-of-the-art sort features, accommodate the excess workload by decreasing sort times, and consequently will also lower the cost per experiment. The BD FACSAria II is requested as a shared instrument to support numerous research projects including, but not limited to, the investigation of: i) RNA interference in lymphocytes;ii) cell cycle control of beta cell mass;iii) the immunobiology of B cell differentiation;iv) COX-2 and PGE2-dependent pathways in lung cancer;v) the development of adrenal cortical cell therapy;vi) carcinogen activation and screening;vii) the M. tuberculosis phagosome;viii) and the molecular regulation of dental stem cell property. The FCCF has established novel staining protocols and standards for sorting of hazardous samples. The FCCF staff will continue to develop and adapt novel technologies for use with the new instrument to support UCLA investigators in their efforts to remain on the cutting edge of science. PUBLIC HEALTH RELEVANCE: We are requesting funding for a highly specialized instrument, a cell sorter, to replace an aging instrument that is not as sensitive, fast, or as good at recovering cells of interest to UCLA biomedical researchers as the cell sorters currently on the market. The UCLA investigators who will use this instrument are attempting to design new therapeutic strategies for inherited disorders, infectious diseases, cancer and aging as well as to develop vaccines and microbicides to prevent infection with pathogens such as HIV. The increased sensitivity of the new instrument will allow these researchers to ask more detailed questions, while the increased speed of the cell sorter will allow more investigators to use it in a single day increasing productivity and decreasing both the cost to the investigator and the time to make important discoveries.